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Cao H, Zhou X, Zeng Y. Microfluidic Exponential Rolling Circle Amplification for Sensitive microRNA Detection Directly from Biological Samples. Sens Actuators B Chem. 2018;279:447-457doi: 10.1016/j.snb.2018.09.121.
Cao, H., Zhou, X., & Zeng, Y. (2019). Microfluidic Exponential Rolling Circle Amplification for Sensitive microRNA Detection Directly from Biological Samples. Sensors and actuators. B, Chemical, 279447-457. https://doi.org/10.1016/j.snb.2018.09.121
Cao, Hongmei, et al. "Microfluidic Exponential Rolling Circle Amplification for Sensitive microRNA Detection Directly from Biological Samples." Sensors and actuators. B, Chemical vol. 279 (2019): 447-457. doi: https://doi.org/10.1016/j.snb.2018.09.121
Cao H, Zhou X, Zeng Y. Microfluidic Exponential Rolling Circle Amplification for Sensitive microRNA Detection Directly from Biological Samples. Sens Actuators B Chem. 2019 Jan 15;279:447-457. doi: 10.1016/j.snb.2018.09.121. Epub 2018 Oct 04. PMID: 30533973; PMCID: PMC6284813.
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Sens Actuators B Chem. 2019 Jan 15;279:447-457. doi: 10.1016/j.snb.2018.09.121. Epub 2018 Oct 04.

Microfluidic Exponential Rolling Circle Amplification for Sensitive microRNA Detection Directly from Biological Samples.

Sensors and actuators. B, Chemical

Hongmei Cao, Xin Zhou, Yong Zeng

Affiliations

  1. Department of Chemistry, University of Kansas, Lawrence, KS 66045.
  2. University of Kansas Cancer Center, Kansas City, KS 66160.

PMID: 30533973 PMCID: PMC6284813 DOI: 10.1016/j.snb.2018.09.121

Abstract

There is an urgent need of sensitive bioanalytical platforms for sensitive and precise quantification of low-abundance microRNA targets in complex biological samples, including liquid biopsies of tumors. Many of current miRNA biosensing methods require laborious sample pretreatment procedures, including extraction of total RNA, which largely limits their biomedical and clinical applications. Herein we developed an integrated Microfluidic Exponential Rolling Circle Amplification (MERCA) platform for sensitive and specific detection of microRNAs directly in minimally processed samples. The MERCA system integrates and streamlines solid-phase miRNA isolation, miRNA-adapter ligation, and a dualphase exponential rolling circle amplification (eRCA) assay in one analytical workflow. By marrying the advantages of microfluidics in leveraging bioassay performance with the high sensitivity of eRCA, our method affords a remarkably low limit of detection at <10 zeptomole levels, with the ability to discriminate single-nucleotide difference. Using the MERCA chip, we demonstrated quantitative detection of miRNAs in total RNA, raw cell lysate, and cellderived exosomes. Comparing with the parallel TaqMan RT-qPCR measurements verified the adaptability of the MERCA system for detection of miRNA biomarkers in complex biological materials. In particular, high sensitivity of our method enables direct detection of low-level exosomal miRNAs in as few as 2 × 10

Keywords: Microfluidic Exponential Rolling Circle Amplification (MERCA); cell-derived exosomes; complex biological samples; microRNA detection; raw cell lysate

Conflict of interest statement

The authors declare no competing financial interest.

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